The invention relates to the technical field of molecular biology and tumor marker medicine, in particular to application of a KLC3 gene as a marker in diagnosis and treatment of lung cancer, gastric cancer, colorectal cancer, endometrial cancer and ovarian cancer. The KLC3 gene can be used as a diagnostic biomarker for lung cancer, gastric cancer, colorectal cancer, endometrial cancer and ovarian cancer or can be used for preparing a cancer diagnosis product. The KLC3 gene is specifically overexpressed in lung cancer, gastric cancer, colorectal cancer, endometrial cancer and ovarian cancer, but there is no such specific high expression in normal tissues. By detecting the expression of the KLC3 gene in a subject, the occurrence of lung cancer, gastric cancer, colorectal cancer, endometrial cancer and ovarian cancer can be quickly, accurately and clearly determined. The KLC3 gene serves as a diagnostic marker of lung cancer, gastric cancer, colorectal cancer, endometrial cancer and ovarian cancer, provides a new target for clinical treatment of lung cancer, gastric cancer, colorectal cancer, endometrial cancer and ovarian cancer, and can be applied to preparation of antitumor drugs.