Fifty-eight percent of voters in the predominantly Italian-speaking canton backed the ‘Ours first' plan initiated by the Swiss People's Party.
The initiative called for companies to give preference to Swiss workers over foreigners if they were equally qualified.
Forty-four percent of voters turned out for the ballot, the result of which puts residents at odds with cantonal leaders.
The Ticino government backed a counter-proposal which suggested the issue should be tackled at federal level as part of Switzerland's ongoing discussions surrounding how to implement immigration quotas without contravening the EU principle of free movement of people.
Eros Sebastiani, the head of a group representing cross border workers, was circumspect after the result.
“We were expecting it. In fact, we thought the percentage would be even higher,” the president of the Associazione Frontalieri Ticino told Italian newspaper La Stampa.
On the other side of the border the vote was greeted with some concern.
Italy's foreign minister, Paolo Gentiloni, played down its immediate importance but said it had the potential to sour relations if pushed through.
"The anti-frontalieri (anti-cross-border workers) referendum has no practical effect for now. But without the free movement of people, Swiss-EU relations would be in jeopardy.”
#Ticino Referendum anti frontalieri non ha per ora effetti pratici Ma senza libera circolazione delle persone rapporti Svizzera-UE a rischio- Paolo Gentiloni (@PaoloGentiloni) September 25, 2016
Lara Comi, the vice president of the European People's Party Group in the European Parliament, was more strident:
“Switzerland doesn't want our cross-border workers. LET'S ISOLATE THEM. Our Ticino-based companies should return to Italy,” wrote the Forza Italia politician.
La Svizzera non vuole I nostri frontalieri.ISOLIAMOLA: nostri imprenditori con sede nel Canton Ticino tornino in Italia @RobertoMaroni_- LARA COMI (@comilara) September 25, 2016
The SVP says it wants to prevent so-called ‘wage-dumping', where foreigners are hired to do a job at a cut-price cost.
Ticino has historically had high numbers of cross-border workers, most coming over from Italy.
Until last year the number of ‘frontalieri' in Ticino rose sharply, with more than a quarter of its labour force living over the Italian border.
Many locals feel that the situation has contributed to raising unemployment and lowering salaries for Swiss workers in the canton.
In 2014, Ticino raised taxes on frontalieri, and in April 2015 authorities imposed tougher conditions on cross-border workers including a requirement that Italian nationals supply a copy of their criminal record.
As a result, figures released earlier this year showed that Ticino was the only canton in Switzerland to see numbers of cross-border workers fall in 2015.
Many politicians in the canton are in agreement with the idea behind the SVP-Ticino's initiative but feel it would not be feasible given it would be limited by current federal and cantonal laws.
The cantonal parliament backed a solution put forward earlier this year by ETH Zurich professor and former minister Michael Ambühl, which suggests temporary limits be put on immigration by region or by job sector should immigration cause problems in those specific areas.
The SVP rejected the counter-proposal, saying their own initiative was a “reflection of the clear NO to the free movement of people”, since Ticino voted overwhelmingly in favour of immigration quotas in the February 9th 2014 referendum.